The Role of XBP1 in bone metabolism.

Bone is a dynamic organ that, once formed, undergoes a constant remodeling process that includes bone resorption and synthesis. Osteoclasts and osteoblasts are primarily responsible for controlling this process. X-box binding protein 1 (XBP1), a transcription factor, affects the metabolism of bones in various ways. In recent years, numerous studies have revealed that XBP1 plays a vital role in bone metabolism, including osteoclast and osteoblast development, as well as in regulating immune cell differentiation that affects the immune microenvironment of bone remodeling. In this review, we highlight the regulatory mechanisms of XBP1 on osteoclasts and osteoblasts, how XBP1 affects the immune microenvironment of bone remodeling by influencing the differentiation of immune cells, and predict the possible future research directions of XBP1 to provide new insights for the treatment of bone-related metabolic diseases.

Exosomal microRNA-223 from neutrophil-like cells inhibits osteogenic differentiation of PDLSCs through the cGMP-PKG signaling pathway.

BACKGROUND AND OBJECTIVE: Neutrophils-derived exosomes have been shown to cause tissue inflammation in many diseases, but their role in periodontitis, a neutrophil-mediated disease, is unknown. Here, we investigated the effect of neutrophil-like cells derived exosomes on osteogenic dysfunction of periodontal ligament stem cells (PDLSCs) in periodontitis. METHODS: Neutrophil-like cells were derived from HL-60 cells by dimethylsulfoxide stimulation. Exosomes were isolated by ultracentrifugation and characterized using transmission electron microscopy, nanoflow cytometry and western blot. MicroRNA-223 (miR-223) expression were analyzed by real-time PCR. Western blot, alkaline phosphatase (ALP), and alizarin red staining were conducted to assess whether exosomes could affect the osteogenic differentiation of PDLSCs. The expression of miR-223 was inhibited in PDLSCs by transfecting with miR-223 inhibitor. Cyclic guanosine monophosphate (cGMP) expression was determined by enzyme-linked immunosorbent assay. RESULTS: We found that miR-223 was significantly increased in neutrophils and neutrophil-like cells derived exosomes. Treatment with exosomes derived from neutrophil-like cells upregulated miR-223 expression and inhibited the osteogenic differentiation of PDLSCs, while transfection with miR-223 inhibitor significantly promoted PDLSCs osteogenic differentiation. In addition, co-treatment with KT5823, a cGMP-PKG pathway inhibitor, markedly abrogated the rescue effects of miR-223 inhibitor on the osteogenic differentiation of PDLSCs. CONCLUSIONS: Our findings suggest that neutrophil-like cells derived exosomes might inhibit osteogenic differentiation of PDLSCs by transporting miR-223 and regulating the cGMP-PKG signaling pathway.

Role of oxidative stress in the relationship between periodontitis and systemic diseases.

Periodontitis is a common inflammatory disease. It is characterized by destruction of the supporting structures of the teeth and could lead to tooth loss and systemic inflammation. Bacteria in inflamed gingival tissue and virulence factors are capable of entering the bloodstream to induce systemic inflammatory response, thus influencing the pathological process of many diseases, such as cardiovascular diseases, diabetes, chronic kidney disease, as well as liver injury. An increasing body of evidence show the complex interplay between oxidative stress and inflammation in disease pathogenesis. When periodontitis occurs, increased reactive oxygen species accumulation leads to oxidative stress. Oxidative stress contributes to major cellular components damage, including DNA, proteins, and lipids. In this article, the focus will be on oxidative stress in periodontal disease, the relationship between periodontitis and systemic inflammation, and the impact of periodontal therapy on oxidative stress parameters.

MicroRNA-223 negatively regulates the osteogenic differentiation of periodontal ligament derived cells by directly targeting growth factor receptors.

BACKGROUND: MicroRNA (miRNA) is accepted as a critical regulator of cell differentiation. However, whether microRNA-223 (miR-223) could affect the osteogenic differentiation of periodontal ligament (PDL)-derived cells is still unknown. The aim of this study was to explore the mechanisms underlying the roles of miR-223 in the osteogenesis of PDL-derived cells in periodontitis. METHODS: Microarray analysis and real-time polymerase chain reaction (RT-PCR) were used to identify difference in miR-223 expression pattern between healthy and inflamed gingival tissue. The target genes of miR-223 were predicted based on Targetscan and selected for enrichment analyses based on Metascape database. The gain-and loss-of-function experiments were performed to discuss roles of miR-223 and growth factor receptor genes in osteogenic differentiation of PDL-derived cells. The target relationship between miR-223 and growth factor receptor genes was confirmed by a dual luciferase assay. Osteogenic differentiation of PDL-derived cells was assessed by Alizarin red staining, RT-PCR and western blot detection of osteogenic markers, including osteocalcin (OCN), osteopontin (OPN) and runt-related transcription factor 2 (Runx2). RESULTS: MiR-223 was significantly increased in inflamed gingival tissues and down-regulated in PDL-derived cells during osteogenesis. The expression of miR-223 in gingival tissues was positively correlated with the clinical parameters in periodontitis patients. Overexpression of miR-223 markedly inhibited PDL-derived cells osteogenesis, which was evidenced by reduced Alizarin red staining and osteogenic markers expressions. Furthermore, two growth factor receptor genes, including fibroblast growth factor receptor 2 (FGFR2) and transforming growth factor beta receptor 2 (TGFßR2), were revealed to be direct targets of miR-223 and shown to undergo up-regulation in PDL-derived cells during osteogenesis. Moreover, suppression of FGFR2 or TGFßR2 dramatically blocked PDL-derived cells osteogenic differentiation. CONCLUSIONS: Our study provides novel evidence that miR-223 can be induced by periodontitis and acts as a negative regulator of PDL-derived cells osteogenesis by targeting two growth factor receptors (TGFßR2 and FGFR2).

Identification of Key Genes and Pathways Associated with Oxidative Stress in Periodontitis.

Methods: The differentially expressed genes (DEGs) were identified using periodontitis-related microarray from the GEO database, and OS-genes were extracted from GeneCards database. The intersection of the OS-genes and the DEGs was considered as oxidative stress-related DEGs (OS-DEGs) in periodontitis. The Pearson correlation and protein-protein interaction analyses were used to screen key OS-genes. Gene set enrichment, functional enrichment, and pathway enrichment analyses were performed in OS-genes. Based on key OS-genes, a risk score model was constructed through logistic regression, receiver operating characteristic curve, and stratified analyses. Results: In total, 74 OS-DEGs were found in periodontitis, including 65 upregulated genes and 9 downregulated genes. Six of them were identified as key OS-genes (CXCR4, SELL, FCGR3B, FCGR2B, PECAM1, and ITGAL) in periodontitis. All the key OS-genes were significantly upregulated and associated with the increased risk of periodontitis. Functional enrichment analysis showed that these genes were mainly associated with leukocyte cell-cell adhesion, phagocytosis, and cellular extravasation. Pathway analysis revealed that these genes were involved in several signaling pathways, such as leukocyte transendothelial migration and osteoclast differentiation. Conclusion: In this study, we screened six key OS-genes that were screened as risk factors of periodontitis. We also identified multiple signaling pathways that might play crucial roles in regulating oxidative stress damage in periodontitis. In the future, more experiments need to be carried out to validate our current findings.

Saliva microbiome changes in thyroid cancer and thyroid nodules patients.

Objective: Thyroid disease has been reported to associate with gut microbiota, but the effects of thyroid cancer and thyroid nodules on the oral microbiota are still largely unknown. This study aimed to identify the variation in salivary microbiota and their potential association with thyroid cancer and thyroid nodules. Methods: We used 16S rRNA high-throughput sequencing to examine the salivary microbiota of thyroid cancer patients (n = 14), thyroid nodules patients (n = 9), and healthy controls (n = 15). Results: The alpha-diversity indices Chao1 and ACE were found to be relatively higher in patients with thyroid cancer and thyroid nodules compared to healthy controls. The beta diversity in both the thyroid cancer and thyroid nodules groups was divergent from the healthy control group. The genera Alloprevotella, Anaeroglobus, Acinetobacter, unclassified Bacteroidales, and unclassified Cyanobacteriales were significantly enriched in the thyroid cancer group compared with the healthy control group. In contrast, the microbiome of the healthy controls was mainly composed of the genera Haemophilus, Lautropia, Allorhizobium Neorhizobium Pararhizobium Rhizobium, Escherichia Shigella, and unclassified Rhodobacteraceae. The thyroid nodules group was dominated by genre uncultured Candidatus Saccharibacteria bacterium, unclassified Clostridiales bacterium feline oral taxon 148, Treponema, unclassified Prevotellaceae, Mobiluncus, and Acholeplasma. In contrast, the genera unclassified Rhodobacteraceae and Aggregatibacter dominated the healthy control group. The study also found that clinical indicators were correlated with the saliva microbiome. Conclusion: The salivary microbiota variation may be connected with thyroid cancer and thyroid nodules.

Effects of non-surgical periodontal therapy on periodontal clinical data in periodontitis patients with rheumatoid arthritis: a meta-analysis.

BACKGROUNDS: To date, there is still no consensus about the clinical efficacy of non-surgical periodontal therapy in rheumatoid arthritis (RA) patients with periodontitis. Therefore, the aim of this study was to summarize clinical data regarding the efficacy of scaling and root planing (SRP) in patients with RA and periodontitis compared to non-RA periodontitis patients. METHODS: We selected randomized controlled trials (RCTs) that compared periodontal clinical data in RA as compared to non-RA periodontitis patients by searching Embase, PubMed and Cochrane Central Register of Controlled Trials and by manually retrieving from the earliest records to March 8, 2021. The overall effect size of plaque index (PI), gingival index (GI), attachment loss (AL), probing depth (PD) and bleeding on probing (BOP) were calculated by either a fixed or random-effect model, and subgroup analyses were conducted according to the different time points of follow-up. Two investigators extracted the data and assess the accuracy of the obtained results with 95% of Confidence Intervals (CI). Cochrane Collaboration's tool was responsible for the evaluation of the literature quality and the inter-study heterogeneity was evaluated by Q test and I2 statistic. Sensitivity analyses were applied for results with heterogeneity. Publication bias was determined by Begg's test, Egger's test and the trim-and-fill method. RESULTS: Seven RCTs including 212 patients eventually met the inclusion criteria for the study. As the primary results, the change of PD was not statistically significant and in the secondary results changes of PI, GI, AL and BOP were also not statistically significant in RA patients with periodontitis compared to non-RA periodontitis patients. In subgroup analysis, a larger BOP reduction at 3 months, PI and AL reduction at 6 months were observed in patients with RA and periodontitis group. The results of sensitivity analyses had no significant effect. No evidence of potential publication bias was tested. There were some limitations due to the small number of eligible RCTs. CONCLUSIONS: SRP is equally effective in RA as compared to non-RA periodontitis patients. It suggests RA does not affect the clinical efficacy of non-surgical periodontal therapy. These results could serve evidence-based practice.

Non-surgical periodontal treatment improves rheumatoid arthritis disease activity: a meta-analysis.

OBJECTIVES: The aim of the meta-analysis was to clarify the efficacy of non-surgical periodontal treatment (NSPT) in improving rheumatoid arthritis (RA) disease activity. METHODS: A systematic literature search was conducted using the PubMed, Embase, and Cochrane databases up to October 2020. A total of nine studies were included for the comparison of RA-related indicator changes between the NSPT group and no treatment (NT) group. Mean differences (MD) and 95% confidence intervals (CI) were calculated for disease activity score (DAS28), erythrocyte sedimentation rate (ESR), tender joint counts (TJC), swollen joint counts (SJC), visual analogical scale (VAS), morning stiffness (MS), rheumatoid factor (RF), C-reactive protein (CRP), tumor necrosis factor α (TNF-α), and interleukin-6 (IL-6). RESULTS: NSPT induced significant reductions of DAS28 (MD: 0.61, 95% CI: 0.37, 0.85, P < 0.001), TJC (MD: 0.65, 95% CI: 0.37, 0.93, P < 0.001), SJC (MD: 0.67, 95% CI: 0.18, 1.17, P = 0.008), VAS (MD: 0.48, 95% CI: 0.08, 0.88, P = 0.02), and CRP (MD: 0.34, 95% CI: 0.07, 0.64, P = 0.01) in RA patients with periodontitis. Other parameters showed a trend toward reduction, but results were not statistically significant. CONCLUSIONS: This meta-analysis indicates that NSPT could improve RA activity as assessed by DAS28, TJC, SJC, VAS, and CRP. CLINICAL RELEVANCE: The results emphasize the effectiveness and need for periodontal diagnosis and periodontal therapy in rheumatoid arthritis patients to reduce disease activity.

Association of TNF-α-308G/A, -238G/A, -863C/A, -1031T/C, -857C/T polymorphisms with periodontitis susceptibility: Evidence from a meta-analysis of 52 studies.

The association between tumor necrosis factor-alpha (TNF-α-308G/A, -238G/A, -863C/A, -1031T/C, and -857C/T) polymorphism and either chronic (CP) or aggressive (AgP) periodontitis susceptibility was conflicting. This meta-analysis aimed to quantitatively estimate the association.A total of 52 studies involving 5519 patients and 7260 controls were identified through a search of multiple electronic databases. Odds ratios (ORs) and their 95% confidence intervals using allele, homozygous, heterozygous, dominant, and recessive genetic models were computed to assess the strength of the association.The TNF-α-308G/A polymorphism was significantly associated with decreased risks of CP (GG vs AA: OR = 0.353, P < .001; GG+GA vs AA: OR = 0.480, P < .001) and AgP (G vs A: OR = 0.651, P < .001; GG vs AA: OR = 0.306, P < .001; GG+GA vs AA: OR = 0.384, P < .001) in Asians. There were no associations between TNF-α-238G/A, -863C/A, -1031T/C, -857C/T polymorphism and susceptibility to AgP. No associations were also found between CP susceptibility and TNF-α-238G/A, -857C/T polymorphism.These findings supported that TNF-α-308G/A polymorphism might be the protective factors of CP and AgP in Asians, and TNF-α-238G/A, -863C/A, -1031T/C, -857C/T polymorphism is not linked to AgP susceptibility.

Interleukin-10 gene polymorphisms and chronic periodontitis susceptibility: Evidence based on 33 studies.

The purpose of the meta-analysis was to investigate the potential association of interleukin-10 (IL-10) polymorphisms with susceptibility to chronic periodontitis (CP). A total of 33 studies involving 3487 cases and 4356 controls were identified through a search of multiple electronic databases (last search was updated on 19 July 2018). Three single nucleotide polymorphisms (SNPs) were included in the meta-analysis: -1082A>G(rs1800896), -819C>T(rs1800871), and -592C>A(rs1800872). Odds ratios (ORs) and their 95% confidence intervals (CIs) using allele, dominant, and recessive genetic models were computed to assess the strength of the association. The -1082A>G(rs1800896) polymorphism was found to be associated with decreased CP risk in both Caucasians and Latinos under the dominant model. The -819C>T(rs1800871) and -592C>A(rs1800872) polymorphisms were both associated with increased CP risk in Latinos under the allele and dominant models. In Asians, no associations were observed for any of the polymorphisms under all comparison models. The present meta-analysis suggests that the -1082A>G(rs1800896) polymorphism might be a protective factor for CP in both Caucasians and Latinos, but the -819C>T(rs1800871) and -592C>A(rs1800872) polymorphisms might contribute to CP pathogenesis in Latinos.

Undergraduate laboratory experiment on determination of total protein content in milk powder by moving reaction boundary titration.

Laboratory exercises focused on protein quantification are frequently conducted in traditional undergraduate biochemistry laboratory curriculum. The laboratory course described here is designed to provide students with experience in measurement of protein content in milk powder by moving reaction boundary titration (MRBT), a new rapid technique for total protein content determination in milk. In addition, this approach is weakly influenced by nonprotein nitrogen reagents such as melamine and urea. The course was done as three weekly laboratory exercises. First, students established a standard curve for milk protein concentration by MRBT method. Then, students investigated the influence of nonprotein nitrogen reagents on MRBT method. Finally, students made a comparison among three different protein quantification methods (MRBT, Biuret, and Kjeldahl method). From the experiments, students grasped the concept and advantages of MRBT and deepened the understanding of protein quantification. This course offer students the opportunity to be exposed to an advanced technique, which may have practical significance to their future study and work in the life science field. © 2018 International Union of Biochemistry and Molecular Biology, 46(6):644-651, 2018.

Association of serum immunoglobulin-G to Porphyromonas gingivalis with acute cerebral infarction in the Chinese population.

BACKGROUND/PURPOSE: There is evidence supporting an association between ischemic stroke and periodontitis in western countries. Differing genetic backgrounds and lifestyles among populations may affect this association. The aim of our study was to determine whether antibody titers to Porphyromonas gingivalis are associated with acute cerebral infarction in the Chinese population. MATERIALS AND METHODS: This case-control study was conducted on 88 acute cerebral infarction patients and 40 healthy control subjects. Serum immunoglobulin-G (IgG) antibody to P. gingivalis was analyzed by enzyme-linked immune sorbent assay. Serum lipids were determined with the automatic biochemical analyzer. Fibrinogen was measured using automated coagulation analyzer. High-sensitivity C-reactive protein (hs-CRP) and interleukin-6 (IL-6) were quantified using commercial ELISA kits. The intima-media thickness of the common carotid arteries (IMT-CCA) was measured by ultrasonography. RESULTS: The results showed that P. gingivalis IgG antibody levels were significantly higher in acute cerebral infarction cases than in healthy controls (mean ± standard deviation, 11.06 ± 1.49 vs. 9.15 ± 1.70, P < 0.001). There were significant correlations of P. gingivalis IgG titer with total cholesterol (r = 0.34, P = 0.001), low-density lipoprotein (r = 0.39, P < 0.001), apolipoprotein-B (r = 0.30, P = 0.004), hs-CRP (r = 0.35, P = 0.001), IL-6 (r = 0.27, P = 0.011), and IMT-CCA (left: r = 0.306, P = 0.004; right: r = 0.241, P = 0.024). CONCLUSION: Antibody titers to P. gingivalis are associated with acute cerebral infarction in the Chinese population.

Di-n-butyl phthalate (DBP) exposure induces oxidative damage in testes of adult rats.

Di-n-butyl phthalate (DBP) is a ubiquitous environmental pollutant, extensively used as a plasticizer in many products including plastics, cosmetics, and medical devices. Some studies have shown that DBP has potential testicular toxicity. However, the mechanism of action of DBP on male reproduction is not clear. The present study was designed to further investigate the potential male reproductive toxicity of DBP . Oxidative stress was assessed in rat testes as an underlying mechanism. Forty SD adult rats were randomly allotted to four groups, and DBP was administered to each group by oral gavage at doses of 0 (control), 100, 250, and 500 mg/kg/d for 2 consecutive weeks. The results indicated that the reproductive toxicity of DBP is dose-dependent. Body and testicular weight was significantly decreased in rats of DBP exposure at a dose of 500 mg/kg/d. Sperm count and motility were significantly decreased at doses of 250 and 500 mg/kg/d. The same two doses significantly inhibited the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and glutathione (GSH) while the level of malondialdehyde (MDA) was significantly increased in testes of rats. Microscopy with hematoxylin and eosin (HE) staining showed that seminiferous tubules atrophy and seminiferous epithelial cells disintegrated and shed in rats of DBP exposure at doses of 500 mg/kg/d. In conclusion, DBP alters the testicular structure and function, at least partly, by inducing oxidative stress in testes of adult rats.

Molecular modeling and 3D-QSAR studies of indolomorphinan derivatives as kappa opioid antagonists.

Molecular modeling and 3D-QSAR studies were performed on 31 indolomorphinan derivatives to evaluate their antagonistic behaviors on kappa opioid receptor and provide information for further modification of this kind of compounds. Best predictions were obtained with CoMFA standard model (q2 = 0.693, N = 4, r2 = 0.900) and CoMSIA combined model (q2 = 0.617, N = 4, r2 = 0.904). Both models were further validated by an external test set of eight compounds with satisfactory predictions: r2 = 0.607 for CoMFA and r2 = 0.701 for CoMSIA. In addition, the 3D structure of human kappa opioid receptor was constructed based on the crystal structure of bovine rhodopsin, and the CoMSIA contour plots were then mapped into the structural model of kappa opioid receptor-GNTI complex to identify key residues, which might account for kappa antagonist potency and selectivity. The roles of nonconserved Glu297 and conserved Lys227 of human kappa opioid receptor were then discussed.